Compositions comprising unsaturated fatty acids and nitric oxide releasing compounds and use thereof for enhancing cognitive and related functions

ABSTRACT

Compositions include one or more unsaturated fatty acids and one or more nitric oxide releasing compounds. Methods for using such compositions can enhance cognitive function, reduce or prevent a decline of social interaction, reduce or prevent age-related behavioral changes, increase trainability, maintain optimal brain function, facilitate learning and memory, reduce memory loss, retard brain aging, prevent or treat strokes, and/or prevent or treat dementia in an animal. Preferably, the compositions are food compositions useful for enhancing cognitive function in humans and companion animals.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part of U.S. patent applicationSer. No. 12/734,013 filed on Apr. 1, 2010, with is a national stageapplication under 35 U.S.C. §371 of PCT/US2008/013870 filed Dec. 19,2008, which claims priority to U.S. Provisional Application Ser. No.61/010,097 filed Jan. 4, 2008 and U.S. Provisional Application Ser. No.61/137,382 filed Jul. 30, 2008, the entire disclosures of which areincorporated herein by reference.

BACKGROUND OF THE INVENTION

Field of the Invention

The present disclosure relates generally to compositions and methods forenhancing cognitive function and particularly to compositions comprisingunsaturated fatty acids and nitric oxide releasing compounds and theiruse for enhancing cognitive function in animals.

Description of Related Art

Aged or aging animals frequently suffer some degree of cognitiveimpairment. Changes, including decline in cognitive function thatprogresses with age, and age-related changes in brain morphology andcerebrovascular function are commonly observed, e.g., brain aging.Age-related or age-associated cognitive impairment may manifest itselfin many ways, e.g., short-term memory loss, diminished capacity tolearn, diminished rate of learning, diminished attention, diminishedmotor performance, and/or dementia, among other indicia. In some cases,a specific etiology of such cognitive decline is unknown. In othercases, cognitive impairment results from the onset or progression ofrecognized diseases, disorders, or syndromes, for example, Alzheimer'sdisease (AD). It is known that age-associated cognitive decline isdistinct from and can occur independently of AD.

Animal models of cognitive impairment greatly facilitate the study ofsuch conditions including their physiology, neurology, anatomy, andpathology, Dogs are useful model animals that demonstrate age-associatedcognitive decline in learning and memory that varies depending on thefunction of the cognitive task (Adams B et al., 2000a; Chan A DF et al.,2002; Su M-Y et al., 1998; and, Tapp P D et al., 2003). While the studyof such decline in dogs is useful in its own right because of their roleas companion animals, the fact that the observed decline mirrorsage-related cognitive declines seen in humans (Adams B et al. 2000b)makes the studies even more valuable. Aged dogs develop neuropathologythat is related to that seen in both successfully aging humans andpatients with AD, such as beta amyloid protein (Cotman C W andBerchtold, 2002; and Cummings B J et al., 1996). However, dogs do notdemonstrate every hallmark of AD, in particular, tau-containingneurofibrillar tangles (Dimakopoulos A C et al., 2002) have not beenobserved. Therefore, the condition in dogs is distinct and referred toas Canine Cognitive Dysfunction Syndrome (CCDS).

Both healthy dogs and unhealthy dogs such as those diagnosed with CCDSmay present clinically with progressive cognitive impairment andneuropathological changes (London E D et al., 1983). In addition, agingdogs and those diagnosed with CCDS exhibit various behavioral disorders.For example, they may not respond to their name or familiar commands,may get lost or confused even in familiar surroundings, may no longergreet or respond to their owners or visitors, may exhibit diminisheddaytime activity, may walk in circles, may shun affection, and may losebladder or bowel control.

Though advances have been made, there remains a need to developcompositions and methods that improve cognition, particularly in aginghumans and other animals. Compositions and methods for the treatmentand/or prevention of cognitive impairment, neurodegeneration conditions,stroke, and dementia are also needed. Such therapies would be useful toimprove the overall quality of life for all involved. For companionanimals, these therapies would lead to improved owner satisfaction andwould improve the owner-companion animal bond.

SUMMARY

It is, therefore, an object of the present disclosure to providecompositions and methods useful for enhancing cognitive function in ananimal.

It is another object of the present disclosure to provide methods forreducing or preventing a decline of social interaction, reducing orpreventing age-related behavioral changes, increasing trainability,maintaining optimal brain function, facilitating learning and memory,reducing memory loss, retarding brain aging, preventing or treatingstrokes, and preventing or treating dementia in an animal.

It is a further object of the present disclosure to provide articles ofmanufacture in the form of kits that contain one or more of thecompounds useful to produce the composition of the present disclosure incombination with foods or other ingredients and devices useful forenhancing cognitive function in an animal.

It is another object of the present disclosure to provide a packagecomprising a composition of the present disclosure and a label affixedto the package indicating the contents of the package and/or thebenefits of administering the composition to an animal.

One or more of these other objects are achieved using novel compositionsand methods for enhancing cognitive function. Generally, thecompositions comprise one or more unsaturated fatty acids and nitricoxide releasing compounds. The methods generally comprise administeringthe compositions in an amount effective for enhancing cognitivefunction, particularly to prevent, reduce and/or delay the age-relatedcognitive-decline, cognitive impairment, neurodegeneration conditions,stroke, and dementia in an animal.

Other and further objects, features, and advantages of the presentdisclosure will be readily apparent to those skilled in the art.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a table setting forth the parameters of an experimental studydisclosed herein as Example 3.

FIG. 2 contains graphs depicting results regarding neurons from theexperimental study disclosed herein as Example 3.

FIG. 3 contains graphs depicting results regarding co-cultures from theexperimental study disclosed herein as Example 3.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

The following abbreviations may be used herein: AA, arachidonic acid;ALA, alpha-linolenic acid; ANOVA, analysis of variance; DHA,docosahexaenoic acid; DPA, docosapentaenoic acid; EPA, eicosapentaenoicacid; LA, linoleic acid; UFA, unsaturated fatty acids (as used hereinUFA refers to one or more such fatty acids); NO, nitric oxide; NORC,nitric oxide releasing compound or compounds; and L-Arg, L-arginine.

The term “animal” means any animal that could benefit from one or moreof the methods of the present disclosure including enhancing cognitivefunction; altering cognitive, motor, or behavioral function; reducing orpreventing a decline of social interaction; reducing or preventingage-related behavioral changes; increasing trainability; maintainingoptimal brain function; facilitating learning and memory; reducingmemory loss; retarding brain aging; preventing or treating strokes;preventing or treating dementia; and/or maintaining mental clarity andalertness. Generally, the animal is a human, avian, bovine, canine,equine, feline, hircine, lupine, murine, ovine, and porcine animal.Preferably, the term “animal” means an animal for which an enhancementof cognitive function is desired or would benefit from an improvement incognitive function. A “companion animal” is any domesticated animal, andincludes, without limitation, cats, dogs, rabbits, guinea pigs, ferrets,hamsters, mice, gerbils, horses, cows, goats, sheep, donkeys, pigs, andthe like. Preferably, the animal is a human or a companion animal suchas a dog or cat.

The term “enhancing cognitive function” means one or more of increasingcognitive, motor, or behavioral function in an animal or preventing,reducing, or delaying a decline in cognitive, motor, or behavioralfunction in an animal.

The term “unsaturated fatty acids” or “UFA” means polyunsaturated fattyacids or monounsaturated fatty acids, including monocarboxylic acidshaving at least one double bond. UFAs include (n-6) fatty acids such aslinoleic acid (LA) and arachidonic acid (AA) and (n-3) fatty acids suchas eicosapentaenoic acid (EPA), alpha-linolenic acid (ALA),docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA). UFAs alsoinclude myristoleic acid, palmitoleic acid, oleic acid, linoleic acid,cis-vaccenic acid, and erucic acid.

The term “fish oil” means a fatty or oily extract, relatively rich inUFA, whether crude or purified, obtained from a sea animal, preferably acold-water fish such as, but not limited to, salmon, tuna, mackerel,herring, sea bass, striped bass, halibut, catfish, and sardines, as wellas shark, shrimp, and clams, or any combination thereof. Fish oil isgenerally a term of art used by ingredient suppliers and encompasses arange or products of varying UFA content and purity.

The term “nitric oxide releasing compounds” or “NORC” means any compoundor compounds that cause or can result in the release of nitric oxide inan animal. Examples of such compounds include L-arginine,L-arginine-containing peptides and proteins, and analogs or derivativesthereof that are known or determined to release nitric oxide, such asarginine alpha-ketoglutarate, GEA 3175, sodium nitroprusside, glyceryltrinitrate, S-nitroso-N-acetyl-penicillamine, nitroglycerin,S—NO-glutathione, NO-conjugated non-steroidal anti-inflammatory drugs(e.g., NO-naproxen, NO-aspirin, NO-ibuprofen, NO-Diclofenac,NO-Flurbiprofen, and NO-Ketoprofen), NO-releasing compound-7,NO-releasing compound-5, NO-releasing compound-12, NO-releasingcompound-18, diazenium diolates and derivatives thereof, diethylamineNONOate, and any organic or inorganic compound, biomolecule, or analog,homolog, conjugate, or derivative thereof that causes the release ofnitric oxide, particularly “free” NO, in an animal. NORC is also definedto include supplements that can be converted to nitric oxide releasingcompounds when metabolized in the body, e.g., citrulline and ornithine.

The term “effective amount” means an amount of a compound, material,composition, medicament, or other material that is effective to achievea particular biological result. Such results include, but are notlimited to, one or more of the following: enhancing cognitive function,increasing daytime activity, improving learning (either the rate or easeof learning), improving attention, improving social behavior, improvingmotor performance, and/or improving cerebrovascular function,particularly in aging animals. In various embodiments, “effectiveamount” refers to an amount suitable to prevent a decline in any one ormore of the above qualities, or, in certain embodiments, to improve anyone or more of the above qualities, for example, cognitive function orperformance, learning rate or ability, problem solving ability,attention span and ability to focus on a task or problem, motor functionor performance, social behavior, and the like. In other embodiments, aneffective amount is suitable to reduce either the extent or rate ofdecline in an animal's cognitive skills or functioning, and/or theeffective amount is suitable to delay the onset of such decline. Sucheffectiveness may be achieved, for example, by administering thecompositions of the present disclosure to an animal or to a populationof animals. Preferably the prevention, reduction, or delay of such adecline, or the improvement in an individual or population is relativeto a cohort, e.g., a control animal or a cohort population that has notreceived the treatment, or been administered the composition ormedicament.

The term “cognitive function” refers to the special, normal, or properphysiologic activity of the brain, including one or more of thefollowing: mental stability, memory/recall abilities, problem solvingabilities, reasoning abilities, thinking abilities, judging abilities,ability to discriminate or make choices, capacity for learning, ease oflearning, perception, intuition, attention, and awareness. “Enhancedcognitive function” or “improved cognitive function” refers to anyimprovement in the special, normal, or proper physiologic activity ofthe brain, including one or more of the following: mental stability,memory/recall abilities, problem solving abilities, reasoning abilities,thinking abilities, judging abilities, ability to discriminate or makechoices, capacity for learning, ease of learning, perception, intuition,attention, and awareness, as measured by any means suitable in the art.

The term “behavior” means anything that an animal does in response orreaction to a given stimulation or set of conditions. “Enhancedbehavior” or “improved behavior” means any improvement in anything thatan animal does in response or reaction to a given stimulation or set ofconditions. “Behavior” is used synonymously with “behavioral function”herein.

The term “motor function” or “motor performance” means the biologicalactivity of the tissues that affect or produce movement in an animal.Such tissue include, without limitation, muscles and motor neurons.“Enhanced motor performance (or function)” or “improved motorperformance (or function)” refers to any improvement in the biologicalactivity of the tissues that affect or produce movement in an animal.

The term “decline” of any of the foregoing categories or specific typesof qualities or functions in an individual (phenotypes) is generally theopposite of an improvement or enhancement in the-quality or function. An“effective amount” of a composition may be an amount required to preventdecline altogether or to substantially prevent decline (“prevent”decline), to reduce the extent or rate of decline (“reduce” decline)over any time course or at any time point, or delay the onset, extent,or progression of a decline (“delay” a decline). Prevention, reduction,or delay of “decline” is frequently a more useful comparative basis whenworking with non-diseased aging animals (e.g., “healthy aging animals”).Prevention, reduction, and delay can be considered relative to a controlor cohort which does not receive the treatment, for example, the diet orsupplement of interest. Prevention, reduction, or delay of either theonset of a detrimental quality or condition, or of the rate of declinein a particular function can be measured and considered on an individualbasis, or in some embodiments on a population basis. The net effect ofpreventing, reducing, or delaying decline is to have less decrease incognitive, motor, or behavioral functioning per unit time, or at a givenend point. In other words, ideally, for an individual or in apopulation, cognitive, motor, and behavioral functioning is maintainedat the highest possible level for the longest possible time. Thus, a netincrease in cognitive, motor, or behavioral function is not required forany embodiment. For purposes herein, an individual can be compared to acontrol individual, group, or population. A population can likewise becompared to an actual individual, to normalized measurements for anindividual, or to a group or population, as is useful.

The term “aging” means being of advanced age such that the animal hasexceeded 50% of the average lifespan for its particular species and/orbreed within a species. For example, if the average lifespan for a givenbreed of dog is 10 years, then a dog within that breed greater than 5years old would be considered “aging” for purposes herein. “Healthyaging animals” are those with no known diseases, particularly diseasesrelating to loss of cognitive impairment such as might confound theresults. In studies using healthy aging animals, cohort animals arepreferably also healthy aging animals, although other healthy animalswith suitable cognitive, motor, or behavioral functioning may besuitable for use as comparative specimens. If animals with specificdisease diagnoses, or cognitive, motor, or behavioral limitations areused, then the cohort animals should include animals that are similarlydiagnosed, or which present with similar indicia of the disease orcognitive, motor, or behavioral limitation.

The term “food” or “food product” or “food composition” means a productor composition that is intended for ingestion by an animal, including ahuman, and provides nutrition to the animal.

As used herein, a “food product formulated for human consumption” is anycomposition specifically intended for ingestion by a human being. Theterm “pet food” or “pet food composition” means a composition intendedfor consumption by animals, preferably by companion animals. A “completeand nutritionally balanced pet food” is one that contains all knownrequired nutrients for the intended recipient or consumer of the food,in appropriate amounts and proportions, based for example onrecommendations of recognized authorities in the field of companionanimal nutrition. Such foods are therefore capable of serving as a solesource of dietary intake to maintain life or promote production, withoutthe addition of supplemental nutritional sources. Nutritionally balancedpet food compositions are widely known and used in the art. The termincludes any food, feed, snack, food supplement, treat, meal substitute,or meal replacement, whether intended for a human or another animal.Animal food includes food or feed intended for any domesticated or wildspecies. In preferred embodiments, a food for an animal represents anutritionally complete food composition, e.g., a pelleted, extruded, ordry food. Examples of such animal foods include extruded pet foods, suchas foods for dogs and cats.

The term “dietary supplement” means a product that is intended to beingested in addition to the normal animal diet. Dietary supplements maybe in any form, e.g., solid, liquid, gel, tablets, capsules, powder, andthe like. Preferably they are provided in convenient dosage forms. Insome embodiments they are provided in bulk consumer packages such asbulk powders, liquids, gels, or oils. In other embodiments, supplementsare provided in bulk quantities to be included in other food items suchas snacks, treats, supplement bars, beverages and the like.

The term “long-term administration” means periods of repeatedadministration or consumption in excess of one month. Periods of longerthan two, three, or four months are preferred for certain embodiments.Also preferred are more extended periods that include longer than 5, 6,7, 8, 9, or 10 months. Periods in excess of 11 months or 1 year are alsopreferred. Longer term use extending over 1, 2, 3, or more years areincluded in the present disclosure. For certain aging animals, theanimal will continue consuming on a regular basis for the remainder ofits life. Sometimes this is referred to as consumption for “extended”periods.

The term “regular basis” means at least monthly dosing with thecompositions or consumption of the compositions, more preferably weeklydosing. More frequent dosing or consumption, such as twice, three, orseven times weekly, is preferred in certain embodiments. Still morepreferred are regimens that comprise at least once daily consumption.The skilled artisan will appreciate that the blood level of a compoundor certain metabolites of that compound or which result after theconsumption of that compound, may be a useful tool for assessing ordetermining dosing frequency. A frequency, regardless of whetherexpressly exemplified herein, that allows maintenance of a desired bloodlevel of the measured compound within acceptable ranges is usefulherein. The skilled artisan will appreciate that dosing frequency willbe a function of the composition that is being consumed or administered,and some compositions may require more or less frequent administrationto maintain a desired blood level of the measured compound.

The term “oral administration” or “orally administering” means that theanimal ingests, or a human is directed to feed, or does feed, the animalone or more of the compositions described herein. Wherein a human isdirected to feed the composition, such direction may be that whichinstructs and/or informs the human that use of the composition mayand/or will provide the referenced benefit, for example, enhancingcognitive function, improving liver function, increasing daytimeactivity, improving learning, improving attention, improving socialbehavior, improving motor performance, and/or improving cerebrovascularfunction, or preventing, reducing, or delaying a decline in suchforegoing functions or qualities. Such direction may be oral direction(e.g., through oral instruction from, for example, a physician,veterinarian, or other health professional, or radio or television media(i.e., advertisement), or written direction (e.g., through writtendirection from, for example, a physician, veterinarian, or other healthprofessional (e.g., prescriptions), sales professional or organization(e.g., through, for example, marketing brochures, pamphlets, or otherinstructive paraphernalia), written media (e.g., internet, electronicmail, website, or other computer-related media), and/or packagingassociated with the composition (e.g., a label present on a containerholding the composition), or a combination thereof (e.g., label orpackage insert with directions to access a website for moreinformation).

The term “cognitive drugs” means any compound, composition, or druguseful for affecting cognitive function, e.g., monoamine oxidase Binhibitors such as selegiline; vasodilators such as nicerogoline andvinpocetine; phosphatidylserine; propentofyline; anticholinesterases(cholinesterase inhibitors) such as tacrine, galantamine, rivastigmine,vinpocetine, donepezil (ARICEPT® (donepezil hydrochloride)),metrifonate, and physostigmine; lecithin; choline cholinomimetics suchas milameline and xanomeline; ionotropic N-methyl-D-aspartate (NMDA)receptor antagonists such as memantine; anti-inflammatory drugs such asprednisolone, diclofenac, indomethacin, propentofyline, naproxen,rofecoxin, ibruprofen and suldinac; metal chelating agents such ascliquinol; Ginkgo biloba; bisphosophonates; selective oestrogen receptormodulators such as raloxifene and estrogen; a phytoestrogen; beta andgamma secretase inhibitors; cholesterol-lowering drugs such as statins;calcitonin; risedronate; alendronate; and combinations thereof.

The term “in conjunction” means that a composition comprising UFA, NORC,a food composition, cognitive drug, or other compound or composition ofthe present disclosure are administered to an animal (1) together in afood composition or (2) separately at the same or different frequencyusing the same or different administration routes at about the same timeor periodically. “Periodically” means that the agent is administered ona dosage schedule acceptable for a specific agent and that the food isfed to an animal routinely as appropriate for the particular animal.“About the same time” generally means that the food and agent areadministered at the same time or within about 72 hours of each other.“In conjunction” specifically includes administration schemes wherein acognitive drug is administered for a prescribed period and thecompositions comprising UFA and NORC are administered indefinitely.

The term “individual” when referring to an animal means an individualanimal of any species or kind.

The term “microorganism” encompasses at least bacteria, molds and otherfungi, and yeasts. Probiotics are beneficial microorganisms that cansurvive or even multiply and thrive in the gastrointestinal tract of ananimal. Probiotics can contribute to the overall health of an animalgenerally and particularly to the gastrointestinal health of the animal.

The term “single package” means that the components of a kit arephysically associated, in or with one or more containers, and considereda unit for manufacture, distribution, sale, or use. Containers include,but are not limited to, bags, boxes or cartons, bottles, packages of anytype or design or material, over-wrap, shrink-wrap, affixed components(e.g., stapled, adhered, or the like), or combinations of any of theforegoing. For example, a single package kit may provide containers ofindividual compositions and/or food compositions physically associatedsuch that they are considered a unit for manufacture, distribution,sale, or use.

The term “virtual package” means that the components of a kit areassociated by directions on one or more physical or virtual kitcomponents instructing the user how to obtain the other components,e.g., in a bag or other container containing one component anddirections instructing the user to go to a website, contact a recordedmessage or a fax-back service, view a visual message, or contact acaregiver or instructor to obtain, for example, instructions on how touse the kit, or safety or technical information about one or morecomponents of a kit. Examples of information that can be provided aspart of a virtual kit include instructions for use; safety informationsuch as material safety data sheets; poison control information;information on potential adverse reactions; clinical study results;dietary information such as food composition or caloric composition;general information on cognitive, behavioral, or motor function;diseases that effect cognitive, behavioral, or motor function; treatingcognitive, behavioral, or motor function; or general information ontreatment or preservation of cognitive, behavioral, or motor function;self-help relating to cognitive, behavioral, or motor function;caregiver information for those caring for animals with cognitive,behavioral, or motor function challenges; and use, benefits, andpotential side-effects or counter-indications for cognitive drugs.

The term “health and wellness of an animal” means the complete physical,mental, and social well being of the animal, not merely the absence ofdisease or infirmity.

The term “extending the prime” means extending the number of years ananimal lives a healthy life and not just extending the number of yearsan animal lives, e.g., an animal would be healthy in the prime of itslife for a relatively longer time.

All percentages expressed herein are by weight of the composition on adry matter basis unless specifically stated otherwise. The skilledartisan will appreciate that the term “dry matter basis” means that aningredient's concentration or percentage in a composition is measured ordetermined after any free moisture in the composition has been removed.

As used throughout, ranges are used herein in shorthand, so as to avoidhaving to set out at length and describe each and every value within therange. Any appropriate value within the range can be selected, whereappropriate, as the upper value, lower value, or the terminus of therange.

As used herein and in the appended claims, the singular form of a wordincludes the plural, and vice versa, unless the context clearly dictatesotherwise. Thus, the references “a”, “an”, and “the” are generallyinclusive of the plurals of the respective terms. For example, referenceto “a puppy”, “a method”, or “a food” includes a plurality of such“puppies”, “methods”, or “foods”. Reference herein, for example to “anantioxidant” includes a plurality of such antioxidants, whereasreference to “pieces” includes a single piece. Similarly, the words“comprise”, “comprises”, and “comprising” are to be interpretedinclusively rather than exclusively. Likewise the terms “include”,“including” and “or” should all be construed to be inclusive, unlesssuch a construction is clearly prohibited from the context. Where usedherein the term “examples,” particularly when followed by a listing ofterms is merely exemplary and illustrative, and should not be deemed tobe exclusive or comprehensive.

The methods and compositions and other advances disclosed here are notlimited to particular methodology, protocols, and reagents describedherein because, as the skilled artisan will appreciate, they may vary.Further, the terminology used herein is for the purpose of describingparticular embodiments only, and is not intended to, and does not, limitthe scope of that which is disclosed or claimed.

Unless defined otherwise, all technical and scientific terms, terms ofart, and acronyms used herein have the meanings commonly understood byone of ordinary skill in the art in the field(s) of the presentdisclosure, or in the field(s) where the term is used. Although anycompositions, methods, articles of manufacture, or other means ormaterials similar or equivalent to those described herein can be used inthe practice of the present disclosure, the preferred compositions,methods, articles of manufacture, or other means or materials aredescribed herein.

All patents, patent applications, publications, technical and/orscholarly articles, and other references cited or referred to herein arein their entirety incorporated herein by reference to the extent allowedby law. The discussion of those references is intended merely tosummarize the assertions made therein. No admission is made that anysuch patents, patent applications, publications or references, or anyportion thereof, are relevant, material, or prior art. The right tochallenge the accuracy and pertinence of any assertion of such patents,patent applications, publications, and other references as relevant,material, or prior art is specifically reserved. Full citations forpublications not cited fully within the specification are set forth atthe end of the specification.

PREFERRED EMBODIMENTS

In one aspect, the present disclosure provides compositions suitable forenhancing cognitive function in an animal. The compositions comprise oneor more unsaturated fatty acids (UFA) and one or more nitric oxidereleasing compounds (NORC) in an amount effective for enhancingcognitive function in an animal. In certain embodiments, thecompositions further comprise one or more B vitamins, one or moreantioxidants, or combinations thereof. The present disclosure is basedupon the discovery that compositions comprising UFA and NORC, with orwithout B vitamins and/or antioxidants, are effective for enhancingcognitive function in animals. The compositions are useful for affectingone or more cognitive, motor, or behavioral functions in animalsregardless of health or age, e.g., juvenile, adult, or senior animals.The compositions increase one or more cognitive, motor, or behavioralfunctions in animals, including healthy animals of all ages or animalsthat are susceptible to or suffering from a decline in cognitivefunction brought about by the aging process or by disease. Similarly,the compositions prevent, reduce, or delay a decline in cognitivefunction in animals, particularly aging animals susceptible to orsuffering from a decline in cognitive function brought about by theaging process or by disease. The compositions are particularly effectivefor reducing or delaying the effects of age-related and disease-relatedcognitive decline in humans and companion animals, particularly dogs andcats. The compositions are also useful for enhancing cognitive functionwhen cognitive decline is caused by changes in brain function,particularly brain aging, or damage from disease, particularly strokes.

The UFA can be any UFA suitable for administration to an animal. UFAscan be obtained from any suitable source, synthetic or natural.Preferred sources of UFA are natural sources of such fatty acids andinclude, without limitation, primrose; dark green vegetables such asspinach; algae and blue-green algae such as spirulina; plant seeds andoils from plants such as flax, canola, soybean, walnut, pumpkin,safflower, sesame, wheat germ, sunflower, corn, and hemp; and fish suchas salmon, tuna, mackerel, herring, sea bass, striped bass, halibut,catfish, sardines, shark, shrimp, and clams; and the extracted oils ofany one or more of the foregoing. The UFA may also be synthetic, and assuch may be produced according to any means suitable in the art, fromany suitable starting material. The UFA may comprise a blend of any oneor more UFA from any one or more sources, such as those exemplifiedabove, whether natural or synthetic. In preferred embodiments, the UFAare long chain polyunsaturated fatty acids (LC-PUFA) comprising one ormore monocarboxylic acids having at least 20 carbon atoms and at leasttwo double bonds. Preferably, the UFAs are (n-6) fatty acids or (n-3)fatty acids, most preferably n-3 LC-PUFAs.

The NORC can be any NORC suitable for administration to an animal. NORCcan be obtained from any suitable source, synthetic or natural. Invarious embodiments, the NORC comprises arginine. Presently preferredsources of arginine include, without limitation, animal and plantproteins. Examples of plants considered rich in arginine content andsuitable for use herein include, but are not limited to, legumes such assoy, lupins, and carob; grains such as wheat and rice; and fruits suchas grapes. Seeds and nuts of plants such as cacao and peanut are alsoconsidered rich in arginine content and are therefore useful herein.Some examples of suitable animal proteins considered rich in argininecontent are poultry and fish products. The NORC can also besynthetically produced, according to any suitable means in the art. Aswith UFA, the NORC content of any composition disclosed herein caninclude a blend of any natural or synthetic NORC. Both UFA and NORC,whether natural or synthetic, can be obtained directly or provided by acommercial source.

In one aspect, the compositions further comprise one or more B vitamins,one or more antioxidants, or combinations thereof. The B vitamins can beany B vitamin suitable for administration to an animal. B vitaminsinclude vitamins B1 (thiamine), B2 (riboflavin), B3 (aka P or PP)(niacin, including nicotinic acid and/or nicotinamide), B5 (pantothenicacid), B6 (pyridoxine), B7 (aka H) (biotin), B8 (myo-inositol), B9 (akaM or B-c) (folic acid), B12 (cobalamin), or salts, conjugates, orderivatives thereof recognized of found to have B vitamin activity.Combinations of any of the foregoing are also useful herein and aresometimes referred to herein as “mixtures” of B vitamins. Since thevitamin requirements vary for different species, not all of the listedcompounds are deemed vitamins for all species. For example, since it isknown that myo-inositol can be synthesized by humans, it is no longerdeemed a vitamin, as it is not required for adequate human nutrition.

The antioxidants can be any antioxidant suitable for administration toan animal. Antioxidants are well known in the art, particularly the artof food technology and food formulation. Natural antioxidant compoundsinclude vitamins (such as A, C and E, and derivative, conjugates, oranalogs thereof), as well as plant extracts, including extracts fromfruit, vegetables, herbs, seeds, and other types and/or parts of plants.Compounds such as -lipoic acid, chlorophyll and derivatives thereof,glutathione, ubiquinols (e.g., coenzyme Q10), carotenoids (e.g.,lycopene), flavonoids, phenolic acids and polyphenols, and pycnogenolare known to be excellent antioxidants. Some examples of plant sourcesof antioxidants include those from fruits such as berries (cherry,blackberry, strawberry, raspberry, crowberry, blueberry, bilberry/wildblueberry, black currant), pomegranate, grape, orange, plum, pineapple,kiwi fruit, and grapefruit; those from vegetables including kale, chilipepper, red cabbage, peppers, parsley, artichoke, Brussels sprouts,spinach, lemon, ginger, garlic, and red beets; those from dry fruitslike apricots, prunes, and dates; from legumes including broad beans,pinto beans, and soybeans. Also nuts and seeds such as pecans, walnuts,hazelnuts, ground nut, and sunflower seeds; cereals such as barley,millet, oats, and corn. Many natural antioxidants are also availablefrom a wide variety of spices including cloves, cinnamon, rosemary, andoregano. Less widely known sources of antioxidants include Ginkgobiloba, and tropical plants such as uyaku, and carica papaya.Antioxidant properties of various teas and green tea, as well asfermented products such as red wine, have become of great interest inrecent years and such would be suitable for use herein. Selenium is anexcellent oxygen scavenger and works well, especially with vitamin orrelated tocopherol compounds. Synthetic dietary antioxidants includebutylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) whichare commonly used in food products. Any of the foregoing, alone or incombination, are suited for use herein, as are combinations of naturaland synthetic antioxidants. In one embodiment, the antioxidants compriseastaxanthin alone or in combination with other antioxidants.

The compositions comprise UFA and NORC, and B vitamins and antioxidantsif included in the composition, in an amount effective for enhancingcognitive function. Generally, the compositions comprise from about 0.1%to about 50% UFA, and from about 0.1% to about 20% NORC. When includedin the composition, the composition comprises from about 0.1 to 40 timesthe recommended daily requirement (RDA) of B vitamins and from about0.0001% to about 25% of antioxidants. In various embodiments, thecompositions comprise from about 1 to about 30% UFA, preferably fromabout 1 to about 15% UFA; and from about 1 to about 15% NORC, preferablyfrom about 1 to about 10% NORC. In various embodiments, the B vitaminscomprise from about 1 to 20 times the RDA, preferably from about 1 to 10times the RDA, and antioxidants comprise from about 0.0001% to about15%, most preferably from about 0.001% to about 5%, most preferably fromabout 0.001% to about 2%. In one embodiment, the compositions compriseUFA, NORC, and a mixture of one or more B vitamins and one or moreantioxidants in such amounts. In one embodiment, the compositioncomprises from about 0.5 g to about 10 g UFA, and from about 0.5 g toabout 10 g NORC, with or without the RDA for B vitamins andantioxidants.

The compositions may further comprise substances such as minerals, othervitamins, salts, functional additives including, for example, palatants,colorants, emulsifiers, antimicrobial or other preservatives. Mineralsthat may be useful in such compositions include, for example, calcium,phosphorous, potassium, sodium, iron, chloride, boron, copper, zinc,magnesium, manganese, iodine, selenium and the like. Examples ofadditional vitamins useful herein include such fat soluble vitamins asA, D, E, and K. Inulin, amino acids, enzymes, coenzymes, and the likemay be useful to include in various embodiments.

In one embodiment, the compositions are food compositions, includinghuman and pet food compositions. Such compositions include foodsintended to supply the necessary dietary requirements for an animal,animal treats (e.g., biscuits), or dietary supplements. The compositionsmay be a dry composition (e.g., kibble), semi-moist composition, wetcomposition, or any mixture thereof. In another embodiment, thecomposition is a dietary supplement such as a gravy, drinking water,beverage, yogurt, powder, granule, paste, suspension, chew, morsel,treat, snack, pellet, pill, capsule, tablet, or any other suitabledelivery form. The dietary supplement can comprise a high concentrationof the UFA and NORC, and optional B vitamins and antioxidants. Thispermits the supplement to be administered to the animal in smallamounts, or in the alternative, can be diluted before administration toan animal. The dietary supplement may require admixing, or preferably beadmixed with water or other diluent prior to administration to theanimal.

In one embodiment, the compositions are refrigerated or frozencompositions. In another embodiment, the UFA and NORC are pre-blendedwith the other components to provide the beneficial amounts needed. Inyet other embodiments, the UFA and NORC are used to coat a food, snack,pet food composition, or pet treat. In one embodiment, the UFA and NORCare added to the composition just prior to offering it to the animal,e.g., using a sprinkled powder or a mix. Such compositions can furthercomprise B vitamins and/or antioxidants.

The compositions can optionally comprise one or more supplementarysubstances that promote or sustain general health. Preferred substancesmay be associated with improved mental health or enhanced cognitivefunction or may be substances that inhibit, delay, or decrease loss ofcognitive function, e.g., herbs or plants that enhance cognitivefunction.

In various embodiments, pet food or pet treat compositions comprise fromabout 15% to about 50% crude protein. The crude protein material maycomprise vegetable proteins such as soybean meal, soy proteinconcentrate, corn gluten meal, wheat gluten, cottonseed, and peanutmeal, or animal proteins such as casein, albumin, and meat protein.Examples of meat protein useful herein include pork, lamb, equine,poultry, fish, and mixtures thereof;

The compositions may further comprise from about 5% to about 40% fat.The compositions may further comprise a source of carbohydrate. Thecompositions may comprise from about 15% to about 60% carbohydrate.Examples of such carbohydrates include grains or cereals such as rice,corn, milo, sorghum, alfalfa, barley, soybeans, canola, oats, wheat, andmixtures thereof. The compositions may also optionally comprise othermaterials such as dried whey and other dairy by-products.

In some embodiments, the ash content of the composition ranges from lessthan 1% to about 15%, preferably from about 5% to about 10%.

The moisture content can vary depending on the nature of thecomposition. In a preferred embodiment, the composition is a completeand nutritionally balanced pet food. In this embodiment, the pet foodmay be a “wet food”, “dry food”, or food of intermediate moisturecontent. “Wet food” describes pet food that is typically sold in cans orfoil bags, and has a moisture content typically in the range of about70% to about 90%. “Dry food” describes pet food which is of a similarcomposition to wet food, but contains a limited moisture content,typically in the range of about 5% to about 15% or 20%, and therefore ispresented, for example, as small biscuit-like kibbles. In one presentlypreferred embodiment, the compositions have moisture content from about5% to about 20%. Dry food products include a variety of foods of variousmoisture contents, such that they are relatively shelf-stable andresistant to microbial or fungal deterioration or contamination. Alsopreferred are dry food compositions which are extruded food products,such as pet foods, or snack foods for either humans or companionanimals.

The compositions may also comprise one or more fiber sources. The term“fiber” includes all sources of “bulk” in the food whether digestible orindigestible, soluble or insoluble, fermentable or nonfermentable.Preferred fibers are from plant sources such as marine plants butmicrobial sources of fiber may also be used. A variety of soluble orinsoluble fibers may be utilized, as will be known to those of ordinaryskill in the art. The fiber source can be beet pulp (from sugar beet),gum arabic, gum talha, psyllium, rice bran, carob bean gum, citrus pulp,fructooligosaccharide, pectin, short chain oligofructose,mannanoligofructose, soy fiber, arabinogalactan, galactooligosaccharide,arabinoxylan, or mixtures thereof.

Alternatively, the fiber source can be a fermentable fiber. Fermentablefiber has previously been described to provide a benefit to the immunesystem of a companion animal. Fermentable fiber or other compositionsknown to skilled artisans that provide a prebiotic to enhance the growthof probiotics within the intestine may also be incorporated into thecomposition to aid in the enhancement of the benefit provided by thepresent disclosure to the immune system of an animal.

In other embodiments, the compositions further comprise prebiotics,probiotics, or a combination thereof. Probiotics are live microorganismsthat have a beneficial effect in the prevention and treatment ofspecific medical conditions when ingested. Probiotics are believed toexert biological effects through a phenomenon known as colonizationresistance. The probiotics facilitate a process whereby the indigenousanaerobic flora limits the concentration of potentially harmful (mostlyaerobic) bacteria in the digestive tract. Other modes of action, such assupplying enzymes or influencing enzyme activity in the gastrointestinaltract, may also account for some of the other functions that have beenattributed to probiotics. Prebiotics are non-digestible food ingredientsthat beneficially affect host health by selectively stimulating thegrowth and/or activity of bacteria in the colon. Prebiotics includefructooligosaccharides (FOS), xylooligosaccharides (XOS),galactooligosaccharides (GOS), and mannooligosaccharides (typically fornon-human foods such as petfoods). The prebiotic, fructooligosaccharide(FOS) is found naturally in many foods such as wheat, onions, bananas,honey, garlic, and leeks. FOS can also be isolated from chicory root orsynthesized enzymatically from sucrose. FOS fermentation in the colonresults in a large number of physiologic effects including increasingthe numbers of bifidobacteria in the colon, increasing calciumabsorption, increasing fecal weight, shortening of gastrointestinaltransit time, and possibly lowering blood lipid levels. The increase inbifidobacteria has been assumed to benefit human health by producingcompounds to inhibit potential pathogens, by reducing blood ammonialevels, and by producing vitamins and digestive enzymes. Probioticbacteria such as Lactobacilli or Bifidobacteria are believed topositively affect the immune response by improving the intestinalmicrobial balance leading to enhanced antibody production and phagocytic(devouring or killing) activity of white blood cells. Bifidobacteriumlactis could be an effective probiotic dietary supplement for enhancingsome aspects of cellular immunity in the elderly. Probiotics enhancesystemic cellular immune responses and may be useful as a dietarysupplement to boost natural immunity in otherwise healthy adults.Probiotics include many types of bacteria but generally are selectedfrom four genera of bacteria: Lactobacilllus acidophillus,Bifidobacteria, Lactococcus and Pediococcus. Beneficial species includeEnterococcus and Saccharomyces species. The amount of probiotics andprebiotics to be administered to the animal is determined by the skilledartisan based upon the type and nature of the prebiotic and probioticand the type and nature of the animal, e.g., the age, weight, generalhealth, sex, extent of microbial depletion, presence of harmfulbacteria, and diet of the animal. Generally, probiotics are administeredto the animal in amounts of from about one to about twenty billioncolony forming units (CFUs) per day for the healthy maintenance ofintestinal microflora, preferably from about 5 billion to about 10billion live bacteria per day. Generally, prebiotics are administered inamounts sufficient to positively stimulate the healthy microflora in thegut and cause these “good” bacteria to reproduce. Typical amounts arefrom about one to about 10 grams per serving or from about 5% to about40% of the recommended daily dietary fiber for an animal. The probioticsand prebiotics can be made part of the composition by any suitablemeans. Generally, the agents are mixed with the composition or appliedto the surface of the composition, e.g., by sprinkling or spraying. Whenthe agents are part of a kit, the agents can be admixed with othermaterials or in their own package.

The compositions and dietary supplements may be specially formulated forthe intended recipients or consumers, such as for adult animals or forolder or young animals. For example, a composition adapted for puppiesor kittens or adapted for active, pregnant, lactating, or aging animalscan be prepared. In general, specialized compositions will compriseenergy and nutritional requirements appropriate for animals at differentstages of development or age.

Certain aspects of the present disclosure are preferably used incombination with a complete and balanced food. According to certainembodiments provided herein, the compositions comprising the UFA andNORC, are preferably used with a high-quality commercial food. As usedherein, “high-quality commercial food” refers to a diet manufactured toproduce the digestibility of the key nutrients of 80% or more, as setforth in, for example, the recommendations of the National ResearchCouncil above for dogs, or in the guidelines set forth by theAssociation of American Feed Control Officials. Similar high nutrientstandards would be used for other animals.

In one embodiment, the food compositions comprise any of a variety ofingredients or combinations thereof selected for their contributions tothe overall composition. Thus a skilled food technologist may choosefrom among natural (e.g., plant or plant-derived, animal, oranimal-derived, and microbial or microbially-derived), and syntheticingredients or components. In particular embodiments, the ingredientsmay include any of the cereal grains and/or fractions or componentsthereof, meat and meat by-products, fish, shellfish, or other seafood,other animal products or by-products, eggs from any source, vitamins,minerals, salts, sweeteners, fiber, flavoring or other palatants,coloring, and functional ingredients such as emulsifiers, stabilizers,softeners, functional coatings, and the like. Cereals useful in theinvention include all plants recognized as “cereal” crops, whethercurrently used in commercial agriculture or merely known practically orbotanically as being a “cereal”. For example, “cereals” includes corn,wheat, rice, barley, sorghum, millet, oats, rye, triticale, buckwheat,fonio, and quinoa. The skilled artisan will appreciate that in a givenfood composition, it is not uncommon to use one or more such cerealproducts. Meats useful in the composition include products from anyanimal, preferably muscle tissue such as chicken or other poultry, lamb,sheep, veal, beef, or pork. Other animal products and by-products usefulin the composition include dairy products or by-products derived fromthe milk of any species. Other important components or ingredientsinclude fats and the skilled artisan will appreciate that many sourcesof vegetable, animal, or microbial fats are available for formulatingfood compositions. In one embodiment, the source of fat is a plant fatsuch as corn, soy, or canola oil, preferably one that is readilyavailable. In another embodiment, an animal fat, such as tallow, isuseful for providing calories from fat, as well as enticing flavor tomeat-eating animals. Of course, combinations of any of the foregoingingredients, such as fats, are known in the art and useful foroptimizing the food compositions based on functional properties as wellas price and availability.

The skilled artisan will also appreciate that in formulating the foodcompositions of the present disclosure, the formulation may varyslightly, so as to allow consideration by the formulator of the priceand/or availability of certain ingredients in the compositions, as wellas the batch-to-batch variation in the analysis of certain ingredients.Thus a given food composition or formulation may vary slightly frombatch to batch, plant to plant, or even season to season depending onsuch factors. Notwithstanding such variation in specific ingredientsselected for manufacturing a particular batch of a food composition, theoverall composition (for example, analysis of protein, carbohydrate,fat, fiber, or other component) may be held constant or at leastsubstantially constant, for example, in accordance with a label claim,such as a claim or guarantee of a minimum or maximum percent of aparticular component.

In other embodiments, the compositions of the present disclosurecomprise UFA and NORC, and, if included in the composition, B vitaminsand antioxidants in an amount effective for one or more of reducing orpreventing a decline of social interaction, reducing or preventingage-related behavioral changes, increasing trainability, maintainingoptimal brain function, facilitating learning and memory, reducingmemory loss, retarding brain aging, preventing or treating strokes,preventing or treating dementia, and maintaining mental clarity andalertness in an animal.

The skilled artisan will understand how to determine the appropriateamount of UFA, NORC, B vitamins, antioxidants, and any other ingredientsto be added to a given composition. The skilled formulator may considerimportant the animal's species, age, size, weight, health, and the likein determining how best to formulate a particular composition, food, orpharmaceutical composition comprising the UFA, NORC, and othercomponents. Other factors that may be taken into account for formulationinclude the type of composition (e.g., pet food composition versusdietary supplement), the desired dosage of each component (UFA, NORC),the average consumption of specific types of compositions by differentanimals (e.g., based on species, body weight, activity/energy demands,and the like) and the manufacturing conditions under which thecomposition is prepared. Preferably, the concentrations of UFA and NORCto be added to the composition are calculated on the basis of the energyand nutrient requirements of the animal. When formulating thecompositions of the present disclosure, a skilled can determine theamounts of the UFA, NORC, and other components of the compositions andof other compounds or ingredients, in for example a food composition,based upon the desired dosages and the characteristics of the animal.

For pet foods and food products formulated for human consumption, theamount of UFA as a percentage of the composition is preferably in therange of about 0.1% to about-13% of the composition, although a greaterpercentage can be supplied. In various embodiments, the amount of UFA isabout 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%,1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%,2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%,3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%,4.8%, 4.9%, 5.0%, or more, e.g., 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%,14%, 15%, 16%, 17%, 18%, 19%, 20% or more, of the composition. Up to 30,40, or 50% UFA may be used in certain embodiments.

For pet foods and food products formulated for human consumption, theamount of NORC as a percentage of the composition is in the range ofabout 0.1% to about 12% of the composition, although a greaterpercentage can be supplied. In various embodiments, the amount of NORCis about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%,1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%,2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%,3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%,4.7%, 4.8%, 4.9%, 5.0%, or more, e.g., 6%, 7%, 8%, 9%, 10%, 11%, 12% ormore, up to about 15% or even 20% of the composition. In specificembodiments, 2-2.5% UFA and 2-2.5% NORC are used. In such embodiments,pure L-arginine is a preferred NORC compound. Dietary supplements may beformulated to contain several-fold higher concentrations of UFA andNORC, to be amenable for administration to an animal in the form of atablet, capsule, liquid concentrate, emulsion, suspension, gel, or otherdosage form, or to be diluted before administration, such as by dilutionin water, or adding to a pet food (for example, by spraying orsprinkling thereon), and other modes of administration suitable foradministering such dietary supplements.

In an alternative embodiment, the amount of. UFA and NORC in thecomposition is a function of an amount required to establish specifiedconcentrations, or a desired range of concentrations, of UFA and/orNORC, or a metabolite thereof, in the blood serum of the animal. Thespecified concentrations, or desired ranges of UFA and/or NORC in theblood serum may be calculated by determining the blood serum levels ofanimals fed the recommended amounts of UFA and NORC specified above, aswould be appreciated by one of skill in the art.

In a preferred embodiment, the food compositions comprise amacronutrient composition suitable for the type of food being designed.In one embodiment, the food composition has about 20 to 32% protein,about 30 to 50% carbohydrate, about 5% to 20% fat, and about 15% to 25%moisture. In another embodiment, the food composition is a pet foodcomposition such as a premium or super-premium pet food composition. Inone embodiment, the pet food is formulated for canines and has a proteincontent of about 20-30%, preferably about 24-28%, and more preferablyabout 25-27%. In one embodiment, the protein content of a dog foodcomposition is about 26% by weight. In another embodiment, theformulation is for felines and has a protein content of about 35-45%,preferably about 37-42%, and more preferably about 39-41%. In oneembodiment, the protein content of a cat food composition is about 40%.In a preferred embodiment, the composition is a food product comprisingUFA and NORC, and further comprising about 15% to about 50% protein,about 5% to about 40% fat, about 5% to about 10% ash content, and havinga moisture content of about 5% to about 20%.

In one embodiment, the food composition is a wet food, such as a cannedfood, frozen food, or fresh food product. In one embodiment, the foodcomposition is shelf stable. In another, it must be refrigerated. Inother embodiments, the food composition is an intermediate moistureproduct or a dry food product as described above.

In a preferred embodiment, the UFA is a fish oil and the NORC isarginine or a nitric oxide-releasing derivative thereof. In certainembodiments, the compositions comprise from about 0.1% to about 50% fishoil and from about 0.1% to about 20% arginine. In preferred embodiments,both UFA and NORC are within the preferred ranges provided herein whenthe composition is a food product. Such ranges offer improvedpalatability and functional properties that will enhance acceptance ofthe food product.

In various embodiments, the composition is a human food composition, petfood composition, or a dietary supplement. The composition that is adietary supplement may contain vastly different concentrations oramounts of the UFA and NORC than a food product or pet food. Generally,the palatability and similar sensory factors are not of concern withcertain dietary supplements, e.g., those that are swallowed.

In one embodiment, the UFA comprises one or more of a natural fish oil,ALA, EPA, DPA, DHA, or another n-3 fatty acid from any source.Combinations of UFA sources are of course contemplated for use herein.UFA with n-3 or n-6 are also contemplated for use in variousembodiments.

Preferably, the composition is formulated for a companion animal, e.g.,a dog or cat. In other embodiments, the animal is a human, with orwithout an age-related cognitive decline.

In one embodiment, the composition is formulated to provide about 0.5 gto about 10 g each of UFA and NORC per day in one or more portions of arecommended serving size. For example, if the composition is intendedfor twice-per-day consumption, a 10 g dose can be provided byformulating the composition such that each recommended portion (e.g., 2spoonsful, or one 20 g portion, or the like) to provide 5 g each of UFAand NORC.

In a preferred embodiment, the compositions comprise from about 0.1% toabout 50% fish oil; and from about 0.1% to about 20% arginine.Preferably, the compositions further comprise from about 0.1 to 40 timesthe recommended daily requirement of B vitamins, from about 0.0001% toabout 25% antioxidants, or both. In one embodiment, the composition usedin the methods comprises from about 0.5 g to about 10 g UFA, and fromabout 0.5 g to about 10 g NORC, with or without B vitamins andantioxidants.

In another aspect, the compositions further comprise one or morecognitive drugs in an amount effective for enhancing cognitive function.The skilled artisan can determine the amount of cognitive drug to beadded to the composition based upon the recommended dosage for the druggiven by its manufacturer or upon the animal's weight, species, age,health status, and the like.

In another aspect, the present disclosure provides pharmaceuticalcompositions comprising a composition of the present disclosure and oneor more pharmaceutically-acceptable carriers, diluents, or excipients.Generally, pharmaceutical compositions are prepared by admixing acompound or composition with excipients, buffers, binders, plasticizers,colorants, diluents, compressing agents, lubricants, flavorants,moistening agents, and the like, including other ingredients known toskilled artisans to be useful for producing pharmaceuticals andformulating compositions that are suitable for administration to ananimal as pharmaceuticals. Optionally, the pharmaceutical compositionsfurther comprise one or both of a B vitamin and an antioxidant.

In another aspect, the present disclosure provides methods for enhancingcognitive function in an animal. The methods generally compriseadministering to an animal a composition comprising one or moreunsaturated fatty acids (UFA) and one or more nitric oxide releasingcompounds (NORC) to the animal in an amount effective for enhancingcognitive function in the animal. In certain embodiments, thecompositions further comprise one or more B vitamins, one or moreantioxidants, or combinations thereof.

In some embodiments, the UFA, NORC, B vitamins, and antioxidants areadministered to the animal in amounts given herein when describing thecompositions. In one embodiment, the composition is administered in anamount effective to increase one or more cognitive, motor, or behavioralfunctions in animals, including animals of all ages that are healthy oranimals that are susceptible to or suffering from a decline in cognitivefunction brought about by the aging process or by disease. In anotherembodiment, the composition is administered in an amount effective toprevent, reduce, or delay a decline in cognitive function in animals,particularly aging animals susceptible to or suffering from a decline incognitive function brought about by the aging process or by disease. Incertain embodiments, the daily dose for the compositions ranges fromabout 5 mg/day to about 5,000 mg/day, 10,000 mg/day, or 20,000, or moreper animal. Preferably, the daily dose ranges from about 30 mg/day toabout 10,000 mg/day per animal, and more preferably from about 750mg/day to about 7,500 mg/day per animal. The daily dose of UFA and NORCcan be measured in terms of grams of UFA and NORC per kg of body weight(BW) of the animal. The daily dose of UFA and NORC, therefore can rangefrom about 0.001 g/kg to about 50 g/kg BW of the animal, althoughgreater or lesser doses can be provided. Preferably, the daily dose ofUFA and NORC is from about 0.001 g/kg to about 25 g/kg BW of the animal.More preferably, the daily dose of UFA and NORC is from about 0.001 g/kgto about 10 g/kg BW of the animal. More preferably, the daily dose ofUFA and NORC is from about 0.001 g/kg to about 5 g/kg BW of the animal.More preferably, the daily dose of UFA and NORC is from about 0.001 g/kgto about 1 g/kg BW of the animal. More preferably, the daily dose of theUFA and NORC is from about 0.001 g/kg to about 0.5 g/kg BW of theanimal.

In another aspect, the present disclosure provides methods for enhancingcognitive function in an animal. The methods generally compriseadministering to an animal a composition comprising one or moreunsaturated fatty acids (UFA) and one or more supplements that can bemetabolized to produce nitric oxide releasing compounds (NORC) to theanimal in an amount effective for enhancing cognitive function in theanimal. In certain embodiments, the compositions further comprise one ormore B vitamins, one or more antioxidants, or combinations thereof. Thesupplements are converted to NORC by the metabolic processes in theanimals and the NORC are involved in enhancing cognitive function asdescribed herein. In preferred embodiments, the supplements arecitrulline/citrulline and ornithine. The supplements can be administeredin any amount sufficient to obtain the NORC amounts required herein uponmetabolism. Typically, the supplements are administered in a compositioncomprising UFA and supplements, preferably about 0.1% to about 50% UFAand 0.1% to about 20% supplement.

Only a portion of the arginine consumed by an individual remainsavailable for metabolization to NO. Arginine is metabolized intocitrulline and NO via the enzyme nitric oxide synthase (NOS). As much as60% of ingested arginine is metabolized in the liver by arginase beforeentering the circulation, where any remaining arginine may bemetabolized to citrulline and NO. An alternative source for arginine isthe endogenous production of arginine from the amino acid citrulline.This route contributes about 20% to whole body arginine production.Citrulline is produced in the intestine and enters the circulationwithout being metabolized by the liver, with almost complete conversionto arginine in the kidneys. Citrulline passes through the liver withlittle or no metabolization and is converted to arginine in themitochondria. The majority of circulating citrulline is converted in thekidneys, which are comprised of highly metabolically active tissue.Citrulline circulating in the bloodstream is first converted to arginineand then in cells to citrulline and NO. Citrulline can be administeredvia the gastrointestinal (GI) tract in oral and enteral products.Citrulline, endogenously, is a product of the metabolism of glutamine inthe gut, generated from ornithine as part of the urea cycle, and formedby nitric oxide synthases distributed in the body. Arginine is generatedfrom citrulline, mainly in the kidney, through its metabolism byargininosuccinate synthase (EC 6.3.4.5) and argininosuccinate lyase (EC4.3.2.1). Significantly, the conversion of citrulline to arginine occurscontinuously, as long as citrulline is circulating in the bloodstream.As a result, circulating citrulline makes it possible to maintainelevated concentrations of arginine over time, which in turn makes itpossible to maintain a steady release of NO in cells. Since the aminoacid citrulline is a precursor to L-arginine, citrulline can substitutefor L-arginine in nutritional compositions. Unlike arginine, citrullineis not metabolized by the liver following entry into the bloodstreamthrough absorption from the diet or de novo intestinal production.Citrulline is enzymatically converted to arginine by mitochondria via apart of the urea cycle.

Similarly, ornithine is a precursor to L-arginine. As a result,ornithine can substitute for L-arginine in nutritional compositions.Unlike arginine, ornithine is not metabolized by the liver followingentry into the bloodstream through absorption from the diet or de novointestinal production. Ornithine is enzymatically converted to arginineby mitochondria via a part of the urea cycle.

Administration in accordance with the methods can be on an as-needed oras-desired basis of varying or regular frequency. A goal of regularingestion is to provide the animal with a regular and consistent dose ofthe composition or the direct or indirect metabolites that result fromsuch ingestion. Such regular and consistent dosing will tend to createconstant blood levels of the components of the compositions or theirdirect or indirect metabolites. Thus, regular administration can be oncemonthly, once weekly, once daily, or more than once daily. Similarly,administration can be every other day, week, or month, every third day,week, or month, every fourth day, week, or month, and the like.Administration can be multiple times per day. When utilized as asupplement to ordinary dietetic requirements, the composition may beadministered directly to the animal, e.g., orally, or otherwise. Thecompositions can alternatively be contacted with, or admixed with, dailyfeed or food, including a fluid, such as drinking water, or anintravenous connection for an animal that is receiving such treatment.When utilized as a daily feed or food, administration will be well knownto those of ordinary skill.

Administration can also be carried out as part of a dietary regimen forthe animal. For example, a dietary regimen may comprise causing theregular ingestion by the animal of a composition described herein in anamount effective to prevent, reduce, or delay a decline in one or morecognitive, motor, or behavioral functions in the animal.

According to the methods of the present disclosure, administration ofthe compositions, including administration as part of a dietary regimen,can span a period of time ranging from parturition through the adultlife of the animal. In various embodiments, the animal is a human orcompanion animal such as a dog or cat. In certain embodiments, theanimal is a young or growing animal. In more preferred embodiments, theanimal is an aging animal. An animal that has reached about 35% of itsprojected lifespan is generally preferred. In presently preferredembodiments administration begins, for example, on a regular or extendedregular basis, when the animal has reached more than about 30%, 40%, or50% of its projected or anticipated lifespan. In some embodiments, theanimal has attained 40, 45, or 50% of its anticipated lifespan. In yetother embodiments, the animal is older having reached 60, 66, 70, 75, or80% of its likely lifespan. A determination of lifespan may be based onactuarial tables, calculations, estimates, or the like, and may considerpast, present, and future influences or factors that are known topositively or negatively affect lifespan. Consideration of species,gender, size, genetic factors, environmental factors and stressors,present and past health status, past and present nutritional status,stressors, and the like may also influence or be taken intoconsideration when determining lifespan.

A composition or method for enhancing cognitive function has manyeffects on an animal. Many day-to-day life functions are dependent uponor related to cognitive function. For example, enhancing cognitivefunction is related to preventing a decline of social interaction,reducing or preventing age-related behavioral changes, increasingtrainability, maintaining optimal brain function, facilitating learningand memory, reducing memory loss, retarding brain aging, preventing ortreating strokes, and preventing or treating dementia in an animal.Similarly, enhancing cognitive function promotes the overall health andwellness of an animal. Further, enhancing cognitive function is integralto extending the prime for an animal. Therefore, enhancing cognitivefunction results in a benefit to the animal in one or more of thesefunctions, e.g., an enhanced cognitive function allows the animal tointeract more desirably with other animals, permits the animal to have abetter memory, and counteract the affects of dementia. Basically,enhancing cognitive function results in an improvement in these andsimilar functions.

In various embodiments of the methods, the composition is a human foodcomposition, pet food composition, or a dietary supplement. In otherembodiments, the composition is a food composition further comprisingabout 15% to about 50% protein, about 5% to about 40% fat, about 5% toabout 10% ash content, and having a moisture content of about 5% toabout 20%.

In certain embodiments, the UFA is a fish oil and the NORC is arginineor a nitric oxide-releasing derivative thereof.

In certain embodiments, the composition being administered comprisesfrom about 0.1% to about 50% fish oil and from about 0.1% to about 20%arginine. For embodiments of the methods, as with the compositionsabove, the UFA comprises one or more of a natural fish oil, ALA, EPA,DPA, DHA, another polyunsaturated fatty acid from any source, orcombinations thereof.

In one embodiment, the composition used in the methods is formulated toprovide about 0.5 g to about 10 g each of UFA and NORC per day via oneor more portions of a recommended serving size. The composition canfurther comprise one or more B vitamins and one or more antioxidants.The B vitamin preferably comprises from about 0.1 to about 40 times theRDA and the antioxidant from about 0.0001% to about 25% of thecomposition. Preferably, the composition comprises a mixture of Bvitamins.

In one embodiment, the composition is administered to the animal inconjunction with one or more cognitive drugs in an amount effective forenhancing cognitive and related functions as defined herein. In aparticular embodiment, the composition administered is thepharmaceutical composition that includes a cognitive drug along with UFAand NORC. In a preferred embodiment, the composition is administered tothe animal on a daily basis, preferably in a single dose.

In certain embodiments, the animal is a healthy aging animal. In others,the animal has a phenotype associated with age-related cognitiveimpairment. For example, when compared to a control animal not havingthe phenotype, the animal may have a phenotype that includes one or moreof decreased ability to recall, short-term memory loss, decreasedlearning rate, decreased capacity for learning, decreased problemsolving skills, decreased attention span, decreased motor performance,increased confusion, or dementia (Alzheimer's in humans or itsequivalent in other animals).

In another aspect, the composition comprises UFA and NORC in an amounteffective for improving one or more social behaviors. In a preferredembodiment, the animal is a companion animal.

The compositions, after being administered to an animal, are believed toincrease the circulating concentrations of UFA and NORC in the blood orblood plasma of the animal. In certain embodiments, the bloodconcentration of a direct or indirect metabolite that results from theconsumption of UFA and/or NORC is increased, and is useful as anindicator of dose. A decrease in concentration of one or more compoundsin the bloodstream of an animal receiving the compositions can alsooccur in the normal course. Such decrease can also be a useful tool formonitoring or determining dosages. Preferably, the change in amount ofthe bloodstream component to be measured is dosage dependent. In someembodiments, the method results in an increase in one or more ketonebodies in the animal's blood. In embodiments where additional compoundsare included in the compositions, e.g., B vitamins or antioxidants,administration results in an increase in circulating concentrations ofthese compounds and is useful as an indication of dose.

The present disclosure also provides other methods for affecting variouscognitive, motor, or behavioral related functions and for affectingvarious physiological functions that relate to cognitive, motor, orbehavioral functions, e.g., social interaction and brain aging.Generally, the methods comprise administering the compositions of thepresent disclosure to an animal in amounts defined herein for thecompositions and methods.

In one aspect, the present disclosure provides methods for reducing orpreventing a decline of social interaction in an animal comprisingadministering a composition comprising UFA and NORC to the animal in anamount effective to reduce or prevent a decline in social interaction.In preferred embodiments, the compositions further comprise one or moreB vitamins, one or more antioxidants, or both in amounts effective forreducing or preventing a decline of social interaction in an animal. Themethods ensure that an aging animal remains involved in playtime,participates in group activities, interacts with caregivers, and thelike.

In one aspect, the present disclosure provides methods for reducing orpreventing a decline of social interaction in an animal comprisingadministering a composition comprising UFA and NORC to the animal in anamount effective to reduce or prevent a decline in social interaction.In preferred embodiments, the compositions further comprise one or moreB vitamins, one or more antioxidants, or both in amounts effective forreducing or preventing a decline of social interaction in an animal. Themethods ensure that an aging animal remains involved in playtime,participates in group activities, interacts with caregivers, and thelike.

In another aspect, the present disclosure provides methods for reducingor preventing age-related behavioral changes in an animal comprisingadministering UFA and NORC to the animal in an amount effective forreducing or preventing age-related behavioral changes in the animal.Preferably, the age-related behavioral changes are one or more offorgetfulness, disorientation, reduced social interaction, changes insleep and wake habits (particularly an increase in nighttime activity),loss of “housetraining” that results in changes in urination anddefecation locations and patterns, confusion, frustration, change intemperament such as agitation and aggression, pacing, and wandering.Such changes have been noted in response to cognitive declines in humansand other animals. In preferred embodiments, the compositions furthercomprise one or more B vitamins, one or more antioxidants, or both inamounts effective for reducing or preventing age-related behavioralchanges in an animal.

In another aspect, the present disclosure provides methods forincreasing trainability of an animal comprising administering acomposition comprising UFA and NORC to the animal in an amount effectivefor increasing trainability. For example, administering the compositionwhile “potty training” babies, puppies, and kittens permits the animalto learn the task more quickly than if the training occurred withoutusing the composition. Similarly, training a dog or cat to obey verbal,signal, or other commands permits the animal to learn the task morequickly than if the training occurred without using the composition.Similarly, the compositions could be useful for training feral or wildanimals such as animals used in a circus or pets raised in the wild. Inpreferred embodiments, the compositions further comprise one or more Bvitamins, one or more antioxidants, or both in amounts effective forincreasing trainability of an animal.

In another aspect, the present disclosure provides methods formaintaining optimal brain function in an animal. The methods compriseadministering a composition comprising UFA and NORC to the animal in anamount effective to prevent or delay a decline in brain function,particularly over time. In preferred embodiments, the compositionsfurther comprise one or more B vitamins, one or more antioxidants, orboth in amounts effective for maintaining optimal brain function in ananimal. Generally, the methods ensure that an aging animal maintainshealthy and optimal brain function throughout life and has a betterquality of life, particularly in older animals. The methods also slowthe progression of mental decline in aging dogs.

In another aspect, the present disclosure provides methods forfacilitating learning and memory in an animal. The methods compriseadministering a composition comprising UFA and NORC to the animal in anamount effective for facilitating learning and memory in the animal. Inone embodiment, the animal has reached at least about 50% of its lifeexpectancy. The methods help an aging animal remember facts andunderstand instructions. In preferred embodiments, the compositionsfurther comprise one or more B vitamins, one or more antioxidants, orboth in amounts effective for facilitating learning and memory in ananimal.

In another aspect, the present disclosure provides methods for reducingmemory loss in an animal. The methods comprise administering acomposition comprising UFA and NORC to the animal in an amount effectiveto reduce memory loss over time. In preferred embodiments, thecompositions further comprise one or more B vitamins, one or moreantioxidants, or both in amounts effective for reducing memory loss inan animal.

In another aspect, the present disclosure provides methods for retardingbrain aging in an animal. The methods comprise administering acomposition comprising UFA and NORC to the animal in an amount effectiveto retard brain aging. In preferred embodiments, the compositionsfurther comprise one or more B vitamins, one or more antioxidants, orboth in amounts effective for retarding brain aging in an animal.

In another aspect, the present disclosure provides methods forpreventing or treating strokes in an animal. The methods compriseadministering a composition comprising UFA and NORC to the animal in anamount effective to prevent or treat strokes. In preferred embodiments,the compositions further comprise one or more B vitamins, one or moreantioxidants, or both in amounts effective for preventing or treatingstrokes in an animal. The methods are based upon the discovery thatreducing damage that results from strokes is correlated to certainaspects of enhancing cognitive function, e.g., reducing memory loss.

In another aspect, the present disclosure provides methods forpreventing or treating dementia in an animal. The methods compriseadministering a composition comprising UFA and NORC to the animal in anamount effective to prevent or treat dementia. In preferred embodiments,the compositions further comprise one or more B vitamins, one or moreantioxidants, or both in amounts effective for preventing or treatingdementia in an animal. Dementia can be Alzheimer's disease (AD) inhumans, Canine Cognitive Dysfunction Syndrome (CCDS) in canines, orsimilar diseases in other animals. The methods are based upon thediscovery that the compositions and methods of the present disclosureprevent or reduce dementia by reducing the effects of damage thatresults from the causes of dementia, e.g., amyloid deposits ordeterioration of artery function.

In another aspect, the present disclosure provides methods formaintaining mental clarity and alertness in an animal. The methodscomprise administering a composition comprising UFA and NORC to theanimal in an amount effective for maintaining mental clarity andalertness. In preferred embodiments, the compositions further compriseone or more B vitamins, one or more antioxidants, or both in amountseffective for maintaining mental clarity and alertness in an animal.

In one aspect, the present disclosure provides methods for promoting thehealth and wellness of an animal comprising administering a compositioncomprising UFA and NORC to the animal in an amount effective to promotehealth and wellness. In preferred embodiments, the compositions furthercomprise one or more B vitamins, one or more antioxidants, or both inamounts effective for promoting health and wellness.

In a further aspect, the present disclosure provides methods forextending the prime for an animal comprising administering a compositioncomprising UFA and NORC to the animal in an amount effective forextending the prime for the animal. In preferred embodiments, thecompositions further comprise one or more B vitamins, one or moreantioxidants, or both in amounts effective for extending the prime forthe animal.

In the methods for reducing or preventing a decline of socialinteraction, reducing or preventing age-related behavioral changes,increasing trainability, maintaining optimal brain function,facilitating learning and memory, reducing memory loss, retarding brainaging, preventing or treating strokes, preventing or treating dementia,maintaining mental clarity and alertness, promoting health and wellness,and extending the prime, the animal is preferably a human or companionanimal, most preferably a dog or cat. The amounts of UFAs, NORC, Bvitamins, antioxidants, and other ingredients used in these methods arethe same as the amounts or within the ranges given for enhancingcognitive function.

In a further aspect, the present disclosure provides kits suitable foradministering a composition comprising one or more UFA and NORC to ananimal. The kits comprise in separate containers in a single package orin separate containers in a virtual package, as appropriate for the kitcomponent, (a) one or more UFAs, (b) one or more NORC, and one or moreof (1) one or more other ingredients suitable for consumption by ananimal; (2) one or more B vitamins; (3) one or more antioxidants; (4)one or more cognitive drugs; (5) one or more prebiotics; (6) one or moreprobiotics; (7) one or more diagnostic devices suitable for determiningwhether an animal could benefit from compositions and methods forenhancing cognitive function and related functions; (8) instructions forhow to combine or prepare the UFA and NORC and any other ingredientsprovided in the kit for administration to an animal; (9) instructionsfor how to use the combined kit components, prepared kit components, orother kit components for-the benefit of an animal; and (10) a device foradministering the combined or prepared kit components to an animal. Thecomponents are each provided in separate containers in a single packageor in mixtures of various components in different packages. In preferredembodiments, the kits comprise the UFAs, NORC, B vitamins, andantioxidants. The kits may comprise the ingredients in variouscombinations. For example, the kit could comprise a mixture of one ormore B vitamins and one or more antioxidants in one container and one ormore other ingredients in one or more other containers. Similarly, thekit could comprise a mixture of UFA and NORC in one container and one ormore other ingredients in one or more other containers. Other suchcombinations can be produced by the skilled artisan based upon thecharacteristics of the ingredients and their physical and chemicalproperties and compatibilities.

In another aspect, the present disclosure provides a means forcommunicating information about or instructions for one or more of (1)using compositions of the present disclosure for enhancing cognitivefunction; (2) admixing the UFAs, NORC, B vitamins, antioxidants, orother components of the present disclosure to produce a compositionsuitable for enhancing cognitive function; (3) using the kits of thepresent disclosure for enhancing cognitive function; and (4)administering the compositions to an animal. The means comprises one ormore of a physical or electronic document, digital storage media,optical storage media, audio presentation, audiovisual display, orvisual display containing the information or instructions. Preferably,the means is selected from the group consisting of a displayed website,a visual display kiosk, a brochure, a product label, a package insert,an advertisement, a handout, a public announcement, an audiotape, avideotape, a DVD, a CD-ROM, a computer readable chip, a computerreadable card, a computer readable disk, a USB device, a FireWiredevice, a computer memory, and any combination thereof

In another aspect, the present disclosure provides methods formanufacturing a food composition comprising UFA, NORC, and one or moreother ingredients suitable for consumption by an animal, e.g., protein,fat, carbohydrate, fiber, B vitamins, and antioxidants. The methodscomprise admixing one or more ingredients suitable for consumption by ananimal with UFA and NORC, and possibly other ingredients such as Bvitamins and/or antioxidants. Alternatively, the methods compriseapplying UFA and NORC, and other ingredients if desired, separately orin any combination onto the food composition, e.g., as a coating ortopping. The UFA and NORC can be added at any time during themanufacture and/or processing of the food composition. This includes,for example, admixing the UFA and NORC as part of the core formulationof the “body” of the food composition or applying them as a coating,i.e., primarily to the surface of the food composition after itsmanufacture. The compositions can be made according to any methodsuitable in the art.

In another aspect, the present disclosure provides a package comprisinga composition of the present disclosure and a label affixed to thepackage containing a word or words, picture, design, acronym, slogan,phrase, or other device, or combination thereof, that indicates that thecontents of the package contains a composition suitable for enhancingcognitive function, reducing or preventing a decline of socialinteraction, reducing or preventing age-related behavioral changes,increasing trainability, maintaining optimal brain function,facilitating learning and memory, reducing memory loss, retarding brainaging, preventing or treating strokes, preventing or treating dementia,or maintaining mental clarity and alertness in an animal, particularlyan aging animal. Typically, such device comprises the words “improvescognitive function”, “improves memory”, “reduces memory loss in aginganimals”, “maintains mental clarity and alertness” or an equivalentexpression printed on the package. Any package or packaging materialsuitable for containing the composition is useful in the presentdisclosure, e.g., a bag, box, bottle, can, pouch, and the likemanufactured from paper, plastic, foil, metal, and the like. In apreferred embodiment, the package contains a food composition adaptedfor a particular animal such as a human, canine or feline, asappropriate for the label, preferably a companion animal foodcomposition.

In another aspect, the present disclosure provides for use of UFA andNORC to prepare a medicament for enhancing cognitive function, reducingor preventing a decline of social interaction, reducing or preventingage-related behavioral changes, increasing trainability, maintainingoptimal brain function, facilitating learning and memory, reducingmemory loss, retarding brain aging, preventing or treating strokes,preventing or treating dementia, and maintaining mental clarity andalertness in an animal. The medicament can further comprise one or moreB vitamins, antioxidants, or combinations thereof. Generally,medicaments are prepared by admixing a compound or composition withexcipients, buffers, binders, plasticizers, colorants, diluents,compressing agents, lubricants, flavorants, moistening agents, and otheringredients known to skilled artisans to be useful for producingmedicaments and formulating medicaments that are suitable foradministration to an animal.

In the present disclosure, the animal can be a juvenile, adult, senior,or geriatric animal. Typically, for most embodiments, the animal is anaging animal. Generally, animals are senior in the last half of theirexpected lifespan and geriatric in the last fourth of their expectedlifespan. Lifespan definitions vary for various animals and are known toskilled artisans. For example, an animal is considered to be juvenileuntil about 16 years of age. For a dog or cat, the animal is consideredto be juvenile until 1 year of age.

The compositions of the present disclosure, including the pharmaceuticalcompositions and medicaments, are administered to the animal using avariety of administration routes. Such routes include oral, intranasal,intravenous, intramuscular, intragastric, transpyloric, subcutaneous,rectal, and the like. Preferably, the compositions are administeredorally.

The compositions of the present disclosure, including the pharmaceuticalcompositions and medicaments, are administered to the animal for a timerequired to accomplish one or more objectives of the present disclosure,e.g., enhancing cognitive function, reducing or preventing a decline ofsocial interaction, reducing or preventing age-related behavioralchanges, increasing trainability, maintaining optimal brain function,facilitating learning and memory, reducing memory loss, retarding brainaging, preventing or treating strokes, preventing or treating dementia,and maintaining mental clarity and alertness in an animal. Thecompositions are suitable for long-term administration or administrationon any schedule compatible with the composition and objective.

In various embodiments of the methods and compositions of the presentdisclosure, the antioxidants comprise astaxanthin alone or incombination with other antioxidants.

EXAMPLES

The present disclosure can be further illustrated by the followingexamples, although it will be understood that the examples are includedmerely for purposes of illustration and are not intended to limit thescope of the present disclosure unless otherwise specifically indicated.

Example 1

Animal and Test Groups

Cognitively-experienced beagle dogs (males and females ranging from 7.5to 11.6 years of age) were tested. The average age of the dogs was 9.38years. To qualify, the dogs must have had at least 6 months of previouscognitive test experience, including having been trained on the delayednon-matching-to position task (DNMP) and the oddity task.

During an initial baseline phase, all dogs were tested on a variabledelay version of the DNMP task (or at 5 s if previously unable to learnthe task) that provided a means of assessing visuo-spatial workingmemory and on a size discrimination and reversal task. Performance onthese tasks was used to place the dogs into four cognitively equivalenttreatment groups (twelve dogs per group): (1) 2% fish oil, 2.5%arginine, 1 to 40 times RDA B vitamins, 150 mg/kg vitamin C, 900 mg/kgvitamin E, and 0.5 mg/kg selenium (“SPB Group”); and (2) the ControlGroup.

Feeding and Water

Dogs had free access to water via wall-mounted automatic wateringsystems and/or water bowls. The animals were provided one of five adultmaintenance foods having approximately 32% protein, 20% fat, and 3%fiber once daily. Initial individual food amounts were calculated usingthe formula “kcal requirement=110 kcal/day×(BW×0.75)”, where BW is bodyweight. The formula was intended to maintain a constant body weight atan appropriate body condition.

Dogs were weighed weekly at the start of treatment and twice monthlyafter food intake and body condition stabilization; food intake wasadjusted as needed to maintain a relatively constant body weight.Animals were given approximately 30 minutes to eat the food provided.The approximate amount of food remaining was recorded in a food intakemonitoring datasheet for each animal.

Housing Conditions

Cognitive Test Battery

Landmark discrimination learning (Days 7-99). Egocentric learning andreversal (Days 100-163).

Landmark Discrimination Task

In this task, the dogs were trained to approach one of two objects,based on their proximity to an external landmark. The task was intendedto assess allocentric spatial ability, which entails utilization ofexternal landmarks to localize objects in space. The dogs were tested onsuccessively more difficult versions of the same general problem.

General test procedures: Dogs were subject to ten (10) trials per day,with an inter-trial interval of 30 seconds. Testing was conducted onceper day, approximately six days per week, such that a total of 80sessions were conducted. A partial correction procedure was used. Inthis procedure, once each session the dogs are permitted to correcttheir response after making an error. Each dog was tested on up to fourproblems, depending upon the dog's success at solving the problems. Tomove on from the first and second problems, the dog had to complete atwo-stage criterion. To pass the first stage, the dog had to respondcorrectly in at least 9 of 10 trials, or on 8 of 10 trials over 2consecutive days. Dogs had to respond in all trials to pass the firststage. The second criterion stage was completed when the dog respondedcorrectly in at least 70% of the next 30 trials over three consecutivesessions. Dogs that had a non-response in any one trial were assigned ascore of 0.5, which was assumed to be the response based on randomchoice, and were given one extra day of testing to complete the 30trials. An average score of 70% over all test days was required to passthe second stage.

The discriminada were identical white coasters and a yellow wooden peg,2 cm×2 cm×9 cm, which served as the landmark. The white circularcoasters were placed over the two lateral food wells on the presentationtray. Velcro tabs 2 cm in diameter, to hold the landmark in place, wereglued to the top center of the coasters and to appropriate loci on thefood tray.

Initial Landmark Test (Land-0): On the initial test (L0), the landmarkwas attached to the center of one of the two white coasters. On eachtrial, the experimenter placed the food reward in either the left orright food well and positioned the landmark accordingly. The door wasraised and the tray was moved to approximately 25 cm from the dog for abrief inspection interval to enable the dog to see the spatialarrangement on the tray. The tray was then presented to the dog and thedog was allowed to respond. In this and all subsequent levels, the dogswere required to respond to the coaster closest to the landmark toobtain the food reward. The correct side was determined randomly by thecomputer with the constraint that each side was correct on half of thetrials of each test session. Each dog was allowed a maximum of 30 testsessions (300 trials) to learn to respond to stimulus associated withthe landmark during L0.

Remedial Training: Dogs that failed the initial L0 test were given aremedial training program to help teach them the task. The remedialtraining consisted of 5 additional training days, with 15 trials perday. At the start of the remedial training, the animals were presentedwith a single rewarded stimulus on the majority of the trials. Withcontinued testing, additional paired stimulus presentations were given.After completing the remedial learning phase, the animals receivedadditional training, up to a maximum of 10 sessions using the originalprotocol. This protocol was also used for dogs that failed L1. Dogs thatfailed L0 were moved on to L1. Dogs that failed remedial training afterL1 were moved on to the next task, if 80 test sessions had not elapsed.If fewer that 10 test sessions remained, the animals on L1 receivedextra remedial training sessions.

Landmark-1 (L1): Once a dog learned the L0 task (either during theinitial training or after the remedial training), the landmark was moved1 cm medially and diagonally away from the edge of the coaster, whichconstituted landmark 1 (L1). The landmark was attached to the food traywith a black piece of 2 cm wide Velcro. Dogs that did not learn within30 sessions were given remedial training, as described above for L0.

Egocentric Test Protocol.

General Test Procedure: At the start of a trial the hinged door wasopened to present the stimulus tray to the dog and the dog was allowed amaximum of 30-sec to make a response by displacing one of the twoidentical stimuli. A correct response was counted when the dog displaceda stimulus object covering food reward. An error was counted when thedog displaced the non-rewarded stimulus. The animals received 12 trialsper session over a period of 63 sessions.

Preference Phase: On the first test day the dogs were given a preferencetest consisting of 10 discrete trials with objects covering both lateralfood wells to determine if any side preferences were present. Thepreferred side was used as the positive side for the initial acquisitionphase of testing. Thus, if the animal chose the object to it's left mostfrequently, then the animals' left side was designated its preferredside. For animals that did not show a side preference (and respondedfive times to each side), a coin toss was used to determine the rewardedside.

Acquisition Phase: Each trial consisted of a single presentation of thestimulus tray with a stimulus covering a reward in the preferred sidelateral well, or the center well. A second non-rewarded stimulus objectcovered a well towards the dog's non-preferred side. Consequently, theobject furthest to the animals' preferred side was always rewarded. Onany given trial, there were three possible spatial configurations(left-center, left, right, or right-center). Each configuration occurredfour times per test session.

Criterion: The acquisition phase was successfully completed once a dogscored a minimum of 90% correct (33/36 correct trials) over threeconsecutive test sessions. A maximum of 12 sessions, or 144 trials, weregiven to meet this criterion. Animals that did not learn within 12sessions were given 5 sessions of remedial training, which was followedby up to 10 additional sessions.

Reversal Phase: Once an animal passed the acquisition phase, therewarded position was switched to the opposite side. Thus, if the objectclosest to a dog's right was rewarded in acquisition testing, the objectclosest to its left was rewarded in the reversal testing. For the firstreversal test, the learning criterion was the same as for the originalacquisition. A maximum of 15 sessions, or 180 trials, were given to meetthis criterion. If an animal did not satisfy the criterion within 15sessions, 5 sessions of remedial training were given. The animals thenreceived 10 additional sessions to reach the learning criterion.

Repeated (Multiple) Reversal Phase: Once an animal learned the originalreversal, the correct side was then be switched and the animal retesteduntil it relearned the task. This procedure was repeated until theanimals had completed a total of 52 sessions on the egocentric protocol.The results are shown in Tables 1 to 5.

Landmark Test Results

TABLE 1 Landmark-0 test results Errors to Criterion Test Group Means SEControl 45.42 12.85 SPB 29.38 8.10

Referring to the data, the SPB Group performed better than the ControlGroup.

TABLE 2 Landmark-1 test results Errors to Criterion Test Group Means SEControl 136.96 16.26 SPB 89.29 19.03

The Control Group differed significantly from the SPB Group.

Egocentric Test Results

TABLE 3 Acquisition test results Errors to Criterion Test Group MeansSEM Control 25.58 2.96 SPB 20.41 2.71

Referring to the data, the Control Group differed from the SPB group.

TABLE 4 Reversal test results Errors to Criterion Test Group Means SEMControl 68.29 6.22 SPB 47.09 5.58

Referring to the data, the Control Group differed significantly from theSPB Group.

TABLE 5 Repeated reversal test results Reversals Learned Test GroupMeans SEM Control 2.4 0.40 SPB 4.42 0.48

Example 2

Male Charles Rivers Sprague-Dawley rats (250 g, Wilmington, Mass.) wereacclimatized to animal facilities 3 days prior to surgery with a 12 hourlight-dark cycle. Bilateral ovariectomy was performed 2 weeks beforediet feeding began. Four weeks after the start of diet feeding,transient middle cerebral artery (tMCA) occlusion under anesthesia wasperformed following intraperitoneal injection of ketamine (60 mgkg) andxylazine (10 mg/kg). 15 rats per group were randomly assigned to one offour treatment groups:

Group 1—Neutered+Control Diet

Group 2—Neutered+Diet II

Group 3—Intact+Control Diet

Group 4—Intact+Diet II

The Control Diet was a standard rat diet containing 140 g/kg casein, 100g/kg sucrose, 50 g/kg fiber, 155 g/kg dextrin, 466 g/kg corn starch, 35g/kg standard salt mix, 40 g/kg soybean oil, 10 g/kg standard vitaminmix, 1.8 g/kg L-cysteine and 2.5 g/kg choline chloride. Diet II was theControl Diet plus 2% Arginine, 2% Menhaden fish oil 4× B vitamins, andantioxidants (Vitamin E: 500 mg/kg diet, Vitamin C: 150 mg/kg diet,Astaxanthin:100 mg/kg, selenium: 0.40 mg/kg).

Animals were anesthetized by an intraperitoneal injection of ketamine(60 mg/kg) and xylazine (10 mg/kg). tMCA occlusion was performed aspreviously described (Simpkins et al., 1997) with slight modification.Briefly, the left common carotid artery, external carotid artery andinternal carotid artery were exposed through a midline cervicalincision. A 3.0 mono-filament suture was introduced into the internalcarotid artery lumen and gently advanced until resistance was felt. Thesurgical procedure was performed within 20 minutes, with minimalbleeding. The suture was kept in place for 60 minutes and then withdrawnto allow reperfusion. Rectal temperature was monitored and maintainedbetween 36.5 and 37° C. with a heating pad throughout the procedure. At24 hours after the onset of reperfusion, the animals were sacrificed andthe brains were removed. The brains were then dissected coronally into 2mm sections using a metallic brain matrix (AS1 Instruments Inc.; Warren,Mich.) and stained by incubation in a 2% solution of2,3,5-triphenyltetrazolium chloride (TTC) in physiological saline at 370C, and then fixed in 10% formalin. DNA fragmentation analysis withTdT-mediated dUTP Nick-End Labeling (TUNEL) MCA occlusion is a widelyused focal ischemic stroke model (Bederson et al., 1986). This in vivomodel for neuronal death can rapidly induce a synchronized apoptoticprocess in a large number of neurons and other cells (Li et al., 1997).The effects of transient ischemia on apoptosis were therefore examinedby analyzing DNA fragmentation with the TUNEL assay. TUNEL staining wasperformed according to the modified manufacturer's instructions(Gavrieli et al., 1992). Formalin-fixed, paraffin-embedded tissuesections were deparaffinizated with xylene, rehydrated through gradedethanol treatment, and given a final wash in PBS. The sections werepost-fixed in 4% paraformaldehyde for 20 minutes. Sections were thenwashed and treated for 15 minutes with 100/pg/ml proteinase K in PBS,equilibrated with equilibration buffer for 10 min, and then incubatedwith buffer containing TdT enzyme and FITC-labeled dUTP (Promega,Madison, Wis.) at 370 C in a humidified chamber. The reaction wasterminated by incubation in 2×SSC buffer for 15 min at room temperature.The sections were then mounted with anti-fade reagents containing DAPI(Molecular Probes, Eugene, Oreg.). Positive control sections wereimmersed in DNase I solution for 10 min at room temperature beforeequilibration in TdT buffer. The sections were observed under afluorescent microscope with appropriate excitation/emission filterpairs. After the surgery, some animals died before the necropsy. Thenumber of animals quantified for lesion volume ranged from 12 to 15 pergroup. The results are shown in Tables 6 and 7.

The results were analyzed with one-way analysis of variance (ANOVA)using Prism software (Graphpad Inc; San Diego, Calif.). The significanceof differences among groups was determined by one-way ANOVA followed byTukey's multiple comparison tests. All values were expressed asmean±SEM.

Referring to Tables 6 and 7, the results show that a diet containing oneor more of the ingredients given in Diet II has a beneficial effect onpreventing or treating ischemia and as such a beneficial effect on manybrain related functions, e.g., cognitive function, social interaction,age-related behavioral changes, decline in brain function, enhancingbrain functions, preventing or treating strokes, retarding brain aging,and preventing or treating dementia such as Alzheimer's disease.

TABLE 6 The Effect of Diet II on the Lesion Volume (mm3) Induced byTransient Focal Cerebral Ischemia Control + Diet II + Intact + Intact +Castration Castration* Control Diet II n 13 12 14 14 Mean 284.9 133.6244.5 163.1 Std. Error 32.50 28.53 43.30 31.54

* p<0.05 Diet II+Castration vs Control+Castration

TABLE 7 The Effect of Diet II on the Apoptotic Index (%) Induced byCerebral Ischemia Control + Diet II + Intact + Intact + CastrationCastration* Control Diet II n 13 12 14 14 Mean 85.55 46.67 75.15 57.69Std. Error 4.134 7.51 8.208 8.137

Example 3

This example provides data from experiments on several neural cellmodels that show the synergistic effect of an NO-releasing compound anda PUFA (e.g., DHA). For a neuronal model, human iCell neurons (CellularDynamics International) were plated in 96-well plates and were grown toneuronal maturity by culturing for 2 weeks. For a co-culture model,human iCell astrocytes and iCell neurons (Cellular DynamicsInternational) were plated in 96-well plates and were grown to neuralmaturity for 2 weeks.

In each model, cells were then treated with DHA (UFA), arginine andcitrulline (NORC's) at 3 different concentrations (see table 1 inFIG. 1) for 2 days. The supernatant was used to perform Amyloid beta 42(A131-42) ELISA Kit, Human (KHB3441, Thermo Fisher Scientific). N=6;One-way ANOVA with Tukey's multiple comparison test, * p<0.05; **p<0.01; *** p<0.001. The results obtained are shown in FIG. 2 and FIG.3.

The release of Aβ1-42 is significantly decreased for the DHA+NORC blendcompared to DHA alone or NORC alone. To this end, it is important tonote that it is known for instance that Aβ1-42 is a major component ofcerebrovascular amyloid deposits and hence this has implications for thepathology of cognitive decline caused by damage from disease such asAlzheimer's disease.

In the left panel of FIG. 2, the combination of UFA+NORC (DHA+arginine)is shown to be significantly different than for the controls (UFA andNORC alone). As shown in the right panel of FIG. 2, citrulline alone hasno effect, nor DHA, but there is a significant decrease for the blend ofUFA+NORC (DHA+citrulline).

FIG. 3 shows that the composition of UFA+NORC is significantly differentthan for the controls (NORC and UFA alone) in the Aβ1-42 test inco-culture.

In view of these experimental results, the observed synergistic effectof a combination of UFA and NORC on neural human cell models whencompared to the control group, provides evidence for the effect of suchcombination on the pathology of cognitive decline caused by damage fromdisease or changes in brain function upon administration of acomposition comprising UFA and NORC.

In the specification, there have been disclosed typical preferredembodiments of the invention. Although specific terms are employed, theyare used in a generic and descriptive sense only and not for purposes oflimitation. The scope of the invention is set forth in the claims.Obviously many modifications and variations of the invention arepossible in light of the above teachings. It is therefore to beunderstood that within the scope of the appended claims the inventionmay be practiced otherwise than as specifically described.

The invention is claimed as follows:
 1. A method for enhancing cognitivefunction in an animal, the method comprising: identifying an animal inwhich enhancement of cognitive function is desired; and administering acomposition comprising effective amounts of one or more unsaturatedfatty acids and one or more nitric oxide releasing compounds on aregular basis, wherein the administration on a regular basis of thecomposition for a time period results in enhanced cognitive function inthe animal compared with an equivalent animal not administered thecomposition for an equivalent time period.
 2. The method of claim 1wherein the animal is an aging animal.
 3. The method of claim 1 whereinthe one or more unsaturated fatty acids comprise one or more of anatural fish oil, ALA, EPA, DPA, DHA or another n-3 fatty acid from anysource.
 4. The method of claim 1 wherein the one or more unsaturatedfatty acids are about 1% to about 15% of the composition.
 5. The methodof claim 1 wherein the one or more nitric oxide releasing compounds areabout 0.1% to about 20% of the composition.
 6. The method of claim 1wherein the composition is administered to the animal with one or morecognitive drugs selected from the group consisting of selegiline,nicerogoline, a phosphatidylserine, propentofyline, galantamine,vinpocetine, donepezil, a Ginkgo biloba extract, a bisphosophonate,raloxifene, an estrogen, a phytoestrogen, calcitonin, risedronate, andalendronate.
 7. The method of claim 1 wherein the composition isadministered to the animal on an extended regular basis.
 8. The methodof claim 1 wherein the composition is administered to the animal on adaily basis.
 9. The method of claim 8 wherein the composition isadministered in a daily dose comprising about 0.001 g of the one or moreunsaturated fatty acids/kg body weight of the animal to about 50 g ofthe one or more unsaturated fatty acids/kg body weight of the animal perday.
 10. The method of claim 8 wherein the composition is administeredin a daily dose comprising about 0.001 g of the one or more unsaturatedfatty acids/kg body weight of the animal to about 0.5 g of the one ormore unsaturated fatty acids/kg body weight of the animal per day. 11.The method of claim 8 wherein the composition is administered in a dailydose comprising about 0.001 g of the one or more nitric oxide releasingcompounds/kg body weight of the animal to about 50 g of the one or morenitric oxide releasing compounds/kg body weight of the animal per day.12. The method of claim 8 wherein the composition is administered in adaily dose comprising about 0.001 g of the one or more nitric oxidereleasing compounds/kg body weight of the animal to about 0.5 g of theone or more nitric oxide releasing compounds/kg body weight of theanimal per day.
 13. The method of claim 1 wherein the animal is ahealthy aging animal, and the administration of the UFA and the NORCresults in enhanced cognitive function in the animal relative to anequivalent healthy aging animal not administered the UFA and the NORC.14. The method of claim 1 wherein the animal has a phenotype associatedwith age-related cognitive impairment, and the administration of the UFAand the NORC results in enhanced cognitive function in the animalrelative to an equivalent animal having the phenotype associated withage-related cognitive impairment and not administered the UFA and theNORC.
 15. The method of claim 14 wherein the phenotype includes one ormore of decreased ability to recall, short-term memory loss, decreasedlearning rate, decreased capacity for learning, decreased problemsolving skills, decreased attention span, decreased motor performance,increased confusion, or dementia, as compared to a control animal nothaving the phenotype.
 16. The method of claim 1 wherein the one or morenitric oxide releasing compounds comprise at least one of arginine,citrulline, or a nitric oxide-releasing derivative of arginine orcitrulline.
 17. The method of claim 16 wherein the one or moreunsaturated fatty acids comprise DHA.
 18. The method of claim 1 whereinthe composition comprises from 0.1 to 40 times the recommended dailyrequirement of B vitamins.
 19. The method of claim 1 wherein thecomposition comprises about 0.0001% to about 25% of antioxidants. 20.The method of claim 1 wherein the animal is a human or a companionanimal.